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Method for detecting histone acetyltransferase based on nano-enzyme

A technology of acetyltransferase and histone, which is applied in the field of nano-biological analysis and detection, can solve the problems that the application of histone acetyltransferase has not been reported, and achieve the effect of low cost, fast response and simple operation

Active Publication Date: 2021-03-23
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its application in the detection of histone acetyltransferases has not been reported

Method used

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  • Method for detecting histone acetyltransferase based on nano-enzyme
  • Method for detecting histone acetyltransferase based on nano-enzyme
  • Method for detecting histone acetyltransferase based on nano-enzyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] a. CuBi 2 o 4 Synthesis of nanomaterials: 1.0mmol Bi(NO 3 ) 3 ·5H 2 O was dispersed in 20mL of aqueous solution containing 0.7mL of concentrated nitric acid, stirred for 2h until the solution was clear and transparent; 10mL of Cu(NO 3 ) 2 ·3H 2 O (0.05mol / L) aqueous solution was added to the above clear solution; then 15mL NaOH aqueous solution (1.0mol / L) was added dropwise under continuous stirring until the solution turned into a blue-green precipitate. The final mixture was transferred to a Teflon-lined stainless steel autoclave and heated at 140 °C for 14 h. The product was alternately washed six times with ethanol and water, and dried under vacuum at 60°C overnight, and the final product was brown powder;

[0038] b. Determination of HAT p300: 20 μL of different concentrations of HAT p300 (0, 0.3, 1.0, 3.0, 10, 30, 100, 150, 200, 300, 500 μmol / L), 20 μL of 1.0 mmol / L acetyl-CoA and 20 μL of 1.0 mmol / L substrate peptide (RGKGGKGLGKGGAKA) were mixed and incub...

Embodiment 2

[0042] a. CuBi 2 o 4 Synthesis of nanomaterials: 1.0mmol BiCl 3 Disperse in 20mL aqueous solution containing 0.7mL concentrated nitric acid, stir for 2h until the solution is clear and transparent; 4 ) 2 ·5H 2 O (0.05mol / L) aqueous solution was added to the above clear solution; then 15mL NaOH aqueous solution (1.0mol / L) was added dropwise under continuous stirring until the solution turned into a blue-green precipitate. The final mixture was transferred to a Teflon-lined stainless steel autoclave and heated at 160 °C for 16 h. The product was alternately washed six times with ethanol and water, and dried under vacuum at 60°C overnight, and the final product was brown powder;

[0043] b. Determination of HAT p300: 20 μL of different concentrations of HAT p300 (0, 0.3, 1.0, 3.0, 10, 30, 100, 150, 200, 300, 500 μmol / L), 20 μL of 1.0 mmol / L acetyl-CoA and 20 μL of 1.0 mmol / L substrate peptide (RGKGGKGLGKGGAKA) were mixed and incubated at 30°C for 80min. After the incubatio...

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PUM

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Abstract

The invention discloses a method for detecting histone acetyltransferase based on nano-enzyme, and belongs to the field of nano-biological analysis and detection. According to the method, CuBi2O4 andK4[Fe(CN)6] undergo an in-situ reaction to generate CuBi2O4 / Bi4 [Fe(CN)6]3 with mimic peroxidase activity, and detection of HAT p300 is performed: when the target compound HAT p300 does not exist, a coenzyme A aptamer is hydrolyzed into dNMP by exonuclease I, and accordingly, generation of the nano-enzyme CuBi2O4 / Bi4 [Fe(CN)6]3 is inhibited; when the HAT p300 exists, the generated coenzyme A and the coenzyme A aptamer are catalyzed to undergo specific recognition, so that the coenzyme A aptamer cannot be hydrolyzed by the exonuclease I, and the CuBi2O4 / Bi4 [Fe(CN)6]3 with the mimic enzyme activity is smoothly generated, and typical chromogenic substrate 3,3',5,5'-tetramethyl benzidine is oxidized, and accordingly, a colorimetric signal is output. The method disclosed by the invention can sensitively detect the HAT p300 within a range of 0.3-200 nmol / L, the detection limit is as low as 0.09 nmol / L, and the method has a very excellent application prospect.

Description

technical field [0001] The invention belongs to the field of nanobiological analysis and detection, and in particular relates to a method for detecting histone acetyltransferase based on nanozyme. Background technique [0002] Histone acetylation is one of the key factors regulating gene transcription and is involved in eukaryotic gene expression and other important life processes as well as other post-translational modifications [Peterson C.L.; Laniel M.A.Curr.Biol.2004,14,R546–R551 ]. Histone acetyltransferase (HAT) acts as a catalytic enzyme for histone acetylation, using acetyl-CoA as an acetyl donor to transfer the acetyl group to the substrate peptide, while producing a large amount of by-product CoA. Evidence has shown that the level of histone acetyltransferase is closely related to the occurrence of various human diseases, such as cancer, leukemia, Alzheimer's disease, AIDS and cardiovascular diseases [Han Y.; Li H.Anal.Chem.2015 , 87, 9179-9185]. Therefore, it i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/48C12Q1/44G01N21/78G01N21/31B01J31/28
CPCC12Q1/48C12Q1/44G01N21/78G01N21/31B01J31/003B01J31/1805B01J31/28G01N2333/91057G01N2333/922B01J2531/842Y02A50/30
Inventor 王光丽陈彦如孙冬雪董玉明
Owner JIANGNAN UNIV
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